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Evaluation of loop-mediated isothermal amplification assay for detection and typing of human papilloma virus 16 and 18 from endocervical samples.
Indian J Med Microbiol. 2019 Apr-Jun; 37(2):241-247.IJ

Abstract

Background

Many human papillomavirus (HPV) types are associated with cervical cancer (CC). Therefore, HPV genotyping has both clinical and epidemiological importance. HPV 16 and 18 are two principal high-risk types responsible for more than 70% of all CC cases. Although several commercial and non-commercial genotyping assays are available, there is a need for a cost-effective and sensitive genotyping method for low- and middle-income countries.

Methods

The study was aimed at evaluation of loop-mediated isothermal amplification (LAMP) assay for HPV genotyping in cervical samples. A total of six primer sets for each HPV type were selected for the assay. The LAMP assay was standardised and validated with HPV control panel. Cervical biopsies were subjected to nested multiplex polymerase chain reaction (NM-PCR; as a part of routine diagnostic workup) and LAMP (HPV 16 and 18) simultaneously.

Results

A total of 225 clinical samples were processed during the study period. The sensitivity of the assay was determined using the 10-fold dilutions of positive controls. Both the HPV 16-LAMP and HPV 18-LAMP assays were shown to detect as low as 10 viral copies per reaction, which is similar to that of NM-PCR. The LAMP assay had a good agreement (new cases; 92%, post-chemoradiotherapy [post-CRT]; 89.1%) with NM-PCR for the detection of both HPV 16 and 18. As compared to histology (new cases; 79.8%, post-CRT; 51.3%), LAMP had better agreement with NM-PCR for detection of HPV from post-CRT cases.

Conclusions

We evaluated the LAMP assay for simultaneous detection and typing of HPV 16 and 18. The assay had good agreement with NM-PCR for detection of both HPV 16 and 18. The LAMP assay is a promising tool for HPV genotyping along with routine cervical cytology, especially in resource-constrained settings.

Authors+Show Affiliations

Department of Microbiology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.Department of Microbiology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.Department of Surgical Oncology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.Department of Microbiology, Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31745026

Citation

Mudhigeti, Nagaraja, et al. "Evaluation of Loop-mediated Isothermal Amplification Assay for Detection and Typing of Human Papilloma Virus 16 and 18 From Endocervical Samples." Indian Journal of Medical Microbiology, vol. 37, no. 2, 2019, pp. 241-247.
Mudhigeti N, Kalawat U, Hulikal N, et al. Evaluation of loop-mediated isothermal amplification assay for detection and typing of human papilloma virus 16 and 18 from endocervical samples. Indian J Med Microbiol. 2019;37(2):241-247.
Mudhigeti, N., Kalawat, U., Hulikal, N., & Kante, M. (2019). Evaluation of loop-mediated isothermal amplification assay for detection and typing of human papilloma virus 16 and 18 from endocervical samples. Indian Journal of Medical Microbiology, 37(2), 241-247. https://doi.org/10.4103/ijmm.IJMM_19_58
Mudhigeti N, et al. Evaluation of Loop-mediated Isothermal Amplification Assay for Detection and Typing of Human Papilloma Virus 16 and 18 From Endocervical Samples. Indian J Med Microbiol. 2019 Apr-Jun;37(2):241-247. PubMed PMID: 31745026.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of loop-mediated isothermal amplification assay for detection and typing of human papilloma virus 16 and 18 from endocervical samples. AU - Mudhigeti,Nagaraja, AU - Kalawat,Usha, AU - Hulikal,Narendra, AU - Kante,Meenakshi, PY - 2019/11/21/entrez PY - 2019/11/21/pubmed PY - 2020/4/18/medline KW - Cervical cancer KW - genotyping assay KW - human papillomavirus KW - loop-mediated isothermal amplification KW - nested multiplex polymerase chain reaction SP - 241 EP - 247 JF - Indian journal of medical microbiology JO - Indian J Med Microbiol VL - 37 IS - 2 N2 - Background: Many human papillomavirus (HPV) types are associated with cervical cancer (CC). Therefore, HPV genotyping has both clinical and epidemiological importance. HPV 16 and 18 are two principal high-risk types responsible for more than 70% of all CC cases. Although several commercial and non-commercial genotyping assays are available, there is a need for a cost-effective and sensitive genotyping method for low- and middle-income countries. Methods: The study was aimed at evaluation of loop-mediated isothermal amplification (LAMP) assay for HPV genotyping in cervical samples. A total of six primer sets for each HPV type were selected for the assay. The LAMP assay was standardised and validated with HPV control panel. Cervical biopsies were subjected to nested multiplex polymerase chain reaction (NM-PCR; as a part of routine diagnostic workup) and LAMP (HPV 16 and 18) simultaneously. Results: A total of 225 clinical samples were processed during the study period. The sensitivity of the assay was determined using the 10-fold dilutions of positive controls. Both the HPV 16-LAMP and HPV 18-LAMP assays were shown to detect as low as 10 viral copies per reaction, which is similar to that of NM-PCR. The LAMP assay had a good agreement (new cases; 92%, post-chemoradiotherapy [post-CRT]; 89.1%) with NM-PCR for the detection of both HPV 16 and 18. As compared to histology (new cases; 79.8%, post-CRT; 51.3%), LAMP had better agreement with NM-PCR for detection of HPV from post-CRT cases. Conclusions: We evaluated the LAMP assay for simultaneous detection and typing of HPV 16 and 18. The assay had good agreement with NM-PCR for detection of both HPV 16 and 18. The LAMP assay is a promising tool for HPV genotyping along with routine cervical cytology, especially in resource-constrained settings. SN - 1998-3646 UR - https://www.unboundmedicine.com/medline/citation/31745026/Evaluation_of_loop_mediated_isothermal_amplification_assay_for_detection_and_typing_of_human_papilloma_virus_16_and_18_from_endocervical_samples_ L2 - http://www.ijmm.org/article.asp?issn=0255-0857;year=2019;volume=37;issue=2;spage=241;epage=247;aulast=Mudhigeti DB - PRIME DP - Unbound Medicine ER -